TaG-EM barcode-based behavioral measurements
A) TaG-EM barcode lines in either a wild-type or norpA background were pooled and tested in a phototaxis assay. After 30 seconds of light exposure, flies in tubes facing the light or dark side of the chamber were collected, DNA was extracted, and TaG-EM barcodes were amplified and sequenced. Barcode abundance values were scaled to the number of flies in each tube and used to calculate a preference index (P.I.). Average P.I. values for four different TaG-EM barcode lines in both the wild-type and norpA backgrounds are shown (n=3 biological replicates, error bars are +/− S.E.M.). B) The same eight lines used for the sequencing-based TaG-EM barcode measurements were independently tested in the phototaxis assay and manually scored videos were used to calculate a P.I. for each genotype. Average P.I. values for each line are shown (n=3 biological replicates, error bars are +/− S.E.M.). C) Flies carrying different TaG-EM barcodes were collected and aged for one to four weeks and then eggs were collected and egg number and viability was manually scored for each line. In parallel the barcoded flies from each timepoint were pooled, and eggs were collected, aged, and DNA was extracted, followed by TaG-EM barcode amplification and sequencing. Average number of viable eggs per female (manual counts) and average barcode abundance are shown both as a bar plot and scatter plot (n=3 biological replicates for 3 barcodes per condition, error bars are +/− S.E.M.).